Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-CaMKII (Thr286) Antibody #3361

Applications Reactivity Sensitivity MW (kDa) Source
W H M R Endogenous 50 Rabbit

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-CaMKII (Thr286) Antibody detects endogenous levels of CaMKII only when phosphorylated at Thr286.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr286 of human CaMKII. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from Neuro2A cells treated with calcium ionophore (A23187, 2 µM) after a 20-minute preincubation with forskolin (30 μM), using Phospho-CaMKII (Thr286) Antibody (upper) or CaMKII Antibody #3362 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from Neuro2A cells, untreated or calf intestinal phosphatase (CIP)-treated, using Phospho-CaMKII (Thr286) Antibody (upper) or CaMKII Antibody #3362 (lower).

Background

CaMKII is an important member of the calcium/calmodulin-activated protein kinase family, functioning in neural synaptic stimulation and T cell receptor signaling (1,2). CaMKII has catalytic and regulatory domains. Ca2+/calmodulin binding to the CaMKII regulatory domain relieves autoinhibition and activates the kinase (3). The activated CaMKII further autophosphorylates at Thr286 to render the kinase constitutively active (3). The threonine phosphorylation state of CaMKII can be regulated through PP1/PKA. PP1 (protein phosphatase 1) dephosphorylates phospho-CaMKII at Thr286. PKA (protein kinase A) prevents phospho-CaMKII (Thr286) dephosphorylation through an inhibitory effect on PP1 (4).

  1. Hughes, K. et al. (2001) J. Biol. Chem. 276, 36008-36013.
  2. Barria, A. et al. (1997) Science 276, 2042-2045.
  3. Barkai, U. et al. (2000) Mol. Endocrinol. 14, 554-563.
  4. Makhinson, M. et al. (1999) J. Neurosci. 19, 2500-2510.
  5. Banno, Y. et al. (2008) J Neurochem 104, 1372-86.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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